{"gene":"WDR64","run_date":"2026-04-28T23:00:23","timeline":{"discoveries":[{"year":2024,"finding":"WDR64 is a testis-specific WD40 protein with two β-propellers that localizes dynamically during spermiogenesis: it is found posterior to the nucleus in steps 8–14 spermatids coinciding with the manchette, moves to the flagella in steps 15–16 spermatids, and localizes at the midpiece of the flagellum in mature spermatozoa.","method":"Immunofluorescence, Western blot, RT-PCR","journal":"Heliyon","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization experiment with functional context, single lab","pmids":["39386799"],"is_preprint":false},{"year":2024,"finding":"WDR64 interacts with ODF1 (outer dense fiber protein 1) to form a protein complex; the WDR64/ODF1 complex co-localizes at the manchette during nuclear shaping and subsequently at the midpiece of mature spermatozoa, implicating it in manchette and flagella assembly during spermiogenesis.","method":"Immunoprecipitation–mass spectrometry (IP-MS), immunofluorescence co-localization","journal":"Heliyon","confidence":"Medium","confidence_rationale":"Tier 2/3 — IP-MS identification of binding partner with supporting co-localization, single lab","pmids":["39386799"],"is_preprint":false}],"current_model":"WDR64 is a testis-specific WD40 repeat protein that dynamically localizes to the manchette and flagellum during spermiogenesis and forms a complex with ODF1, suggesting a role in manchette-mediated nuclear shaping and flagellar assembly."},"narrative":{"teleology":[{"year":2024,"claim":"The first characterization of WDR64 established it as a testis-specific WD40 protein with a dynamic localization pattern during spermiogenesis—moving from the manchette to the flagellum—and identified ODF1 as a physical binding partner, linking WDR64 to nuclear shaping and flagellar assembly.","evidence":"Immunofluorescence, RT-PCR, Western blot, and IP-MS in mouse testis and spermatozoa (single lab study)","pmids":["39386799"],"confidence":"Medium","gaps":["No loss-of-function (knockout or knockdown) data demonstrating a requirement for WDR64 in spermiogenesis or fertility","WDR64–ODF1 interaction identified by IP-MS without reciprocal Co-IP or in vitro reconstitution","Structural basis for the two β-propeller domains and their individual contributions to localization or interaction is unknown"]},{"year":null,"claim":"Whether WDR64 is functionally required for sperm morphogenesis and male fertility remains untested, and the molecular mechanism by which the WDR64–ODF1 complex contributes to manchette-mediated nuclear shaping or flagellar midpiece assembly is unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No genetic studies in animal models or human cohorts linking WDR64 mutations to infertility or spermiogenesis defects","Downstream effectors or cargo transported by the WDR64–ODF1 complex are unidentified","Regulatory mechanisms controlling WDR64 expression timing and dynamic relocalization are unknown"]}],"mechanism_profile":{"molecular_activity":[],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[0,1]}],"pathway":[{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[0,1]}],"complexes":[],"partners":["ODF1"],"other_free_text":[]},"mechanistic_narrative":"WDR64 is a testis-specific WD40 repeat protein containing two β-propellers that dynamically localizes during spermiogenesis, associating with the manchette in elongating spermatids (steps 8–14) and subsequently relocating to the flagellum (steps 15–16) and the midpiece of mature spermatozoa [PMID:39386799]. WDR64 forms a complex with the outer dense fiber protein ODF1, co-localizing at both the manchette during nuclear shaping and the midpiece of mature sperm, implicating it in manchette-mediated nuclear remodeling and flagellar assembly [PMID:39386799]."},"prefetch_data":{"uniprot":{"accession":"B1ANS9","full_name":"WD repeat-containing protein 64","aliases":[],"length_aa":1081,"mass_kda":123.6,"function":"","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/B1ANS9/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/WDR64","classification":"Not Classified","n_dependent_lines":7,"n_total_lines":74,"dependency_fraction":0.0945945945945946},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/WDR64","total_profiled":1310},"omim":[{"mim_id":"621432","title":"WD REPEAT-CONTAINING PROTEIN 64; WDR64","url":"https://www.omim.org/entry/621432"},{"mim_id":"182878","title":"OUTER DENSE FIBER OF SPERM TAILS 1; ODF1","url":"https://www.omim.org/entry/182878"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Not detected","tissue_distribution":"Not detected","driving_tissues":[],"url":"https://www.proteinatlas.org/search/WDR64"},"hgnc":{"alias_symbol":["FLJ32978"],"prev_symbol":[]},"alphafold":{"accession":"B1ANS9","domains":[{"cath_id":"-","chopping":"2-96","consensus_level":"high","plddt":72.7459,"start":2,"end":96},{"cath_id":"2.130.10.10","chopping":"132-248_259-380","consensus_level":"medium","plddt":84.8615,"start":132,"end":380}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/B1ANS9","model_url":"https://alphafold.ebi.ac.uk/files/AF-B1ANS9-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-B1ANS9-F1-predicted_aligned_error_v6.png","plddt_mean":70.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=WDR64","jax_strain_url":"https://www.jax.org/strain/search?query=WDR64"},"sequence":{"accession":"B1ANS9","fasta_url":"https://rest.uniprot.org/uniprotkb/B1ANS9.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/B1ANS9/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/B1ANS9"}},"corpus_meta":[{"pmid":"39386799","id":"PMC_39386799","title":"WDR64, a testis-specific protein, is involved in the manchette and flagellum formation by interacting with ODF1.","date":"2024","source":"Heliyon","url":"https://pubmed.ncbi.nlm.nih.gov/39386799","citation_count":2,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"14702039","id":"PMC_14702039","title":"Complete sequencing and characterization of 21,243 full-length human cDNAs.","date":"2003","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/14702039","citation_count":754,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"20923822","id":"PMC_20923822","title":"Radiation pharmacogenomics: a genome-wide association approach to identify radiation response biomarkers using human lymphoblastoid cell lines.","date":"2010","source":"Genome research","url":"https://pubmed.ncbi.nlm.nih.gov/20923822","citation_count":119,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"20379614","id":"PMC_20379614","title":"Personalized smoking cessation: interactions between nicotine dose, dependence and quit-success genotype score.","date":"2010","source":"Molecular medicine (Cambridge, Mass.)","url":"https://pubmed.ncbi.nlm.nih.gov/20379614","citation_count":108,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"23942779","id":"PMC_23942779","title":"A genome-wide association study of behavioral disinhibition.","date":"2013","source":"Behavior genetics","url":"https://pubmed.ncbi.nlm.nih.gov/23942779","citation_count":91,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"35256949","id":"PMC_35256949","title":"Histone deacetylase inhibitors inhibit cervical cancer growth through Parkin acetylation-mediated mitophagy.","date":"2021","source":"Acta pharmaceutica Sinica. 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B","url":"https://pubmed.ncbi.nlm.nih.gov/39807310","citation_count":5,"is_preprint":false,"source_track":"gene2pubmed"}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":2010,"output_tokens":347,"usd":0.005618},"stage2":{"model":"claude-opus-4-6","input_tokens":3498,"output_tokens":695,"usd":0.052297},"total_usd":0.131077,"stage1_batch_id":"msgbatch_01BparvgRZ9S2LMidvaoo86K","stage2_batch_id":"msgbatch_01NkBfv9SVZWthhS7j6mU6G3","note":"batch pricing = 50% of standard","round2_stage1":{"model":"claude-sonnet-4-6","input_tokens":6909,"output_tokens":502,"usd":0.014129},"round2_rules_fired":"R3","round2_stage2":{"model":"claude-opus-4-6","input_tokens":3916,"output_tokens":791,"usd":0.059033}},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2024,\n      \"finding\": \"WDR64 is a testis-specific WD40 protein that interacts with ODF1 to form a complex, localizing at the manchette during nucleus shaping and at the midpiece of the mature spermatozoa flagellum, implicating the WDR64/ODF1 complex in manchette and flagella assembly during spermiogenesis.\",\n      \"method\": \"Immunoprecipitation–mass spectrometry (IP–MS) for interaction identification; immunofluorescence for subcellular localization; RT-PCR and Western blot for expression characterization\",\n      \"journal\": \"Heliyon\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single lab, Co-IP/MS interaction with localization data supporting functional context, but no loss-of-function or in vitro reconstitution\",\n      \"pmids\": [\"39386799\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WDR64 is a testis-specific WD40-domain protein that forms a complex with ODF1, dynamically localizing at the manchette during spermatid nucleus shaping and subsequently at the flagellar midpiece, suggesting a role in manchette-mediated intramanchette transport and flagella assembly during spermiogenesis.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2024,\n      \"finding\": \"WDR64 is a testis-specific WD40 protein with two β-propellers that localizes dynamically during spermiogenesis: it is found posterior to the nucleus in steps 8–14 spermatids coinciding with the manchette, moves to the flagella in steps 15–16 spermatids, and localizes at the midpiece of the flagellum in mature spermatozoa.\",\n      \"method\": \"Immunofluorescence, Western blot, RT-PCR\",\n      \"journal\": \"Heliyon\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiment with functional context, single lab\",\n      \"pmids\": [\"39386799\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"WDR64 interacts with ODF1 (outer dense fiber protein 1) to form a protein complex; the WDR64/ODF1 complex co-localizes at the manchette during nuclear shaping and subsequently at the midpiece of mature spermatozoa, implicating it in manchette and flagella assembly during spermiogenesis.\",\n      \"method\": \"Immunoprecipitation–mass spectrometry (IP-MS), immunofluorescence co-localization\",\n      \"journal\": \"Heliyon\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2/3 — IP-MS identification of binding partner with supporting co-localization, single lab\",\n      \"pmids\": [\"39386799\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WDR64 is a testis-specific WD40 repeat protein that dynamically localizes to the manchette and flagellum during spermiogenesis and forms a complex with ODF1, suggesting a role in manchette-mediated nuclear shaping and flagellar assembly.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"WDR64 is a testis-specific WD40-domain protein that interacts with ODF1 and localizes dynamically at the manchette during spermatid nucleus shaping and subsequently at the midpiece of the mature spermatozoon flagellum, implicating the WDR64/ODF1 complex in manchette-mediated intracellular transport and flagella assembly during spermiogenesis [PMID:39386799]. The detailed molecular mechanism of WDR64 function remains uncharacterized, as no loss-of-function or reconstitution studies have been reported.\",\n  \"teleology\": [\n    {\n      \"year\": 2024,\n      \"claim\": \"Identifying WDR64 as a testis-specific WD40 protein that forms a complex with ODF1 and localizes to the manchette and flagellar midpiece established a candidate molecular player in spermatid remodeling and flagella assembly.\",\n      \"evidence\": \"IP–MS interaction mapping, immunofluorescence localization, RT-PCR and Western blot expression profiling in mouse testis\",\n      \"pmids\": [\"39386799\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No loss-of-function (knockout or knockdown) data to confirm a requirement for WDR64 in spermiogenesis\",\n        \"Interaction with ODF1 demonstrated by Co-IP/MS only; no reciprocal pull-down or in vitro reconstitution reported\",\n        \"Molecular activity of the WD40 domain and direct substrates or cargo are unknown\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown whether WDR64 loss causes male infertility, what cargo or signaling pathways WDR64 mediates at the manchette, and whether the WDR64/ODF1 complex interacts with other manchette transport or outer dense fiber components.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No genetic or phenotypic data linking WDR64 to infertility or spermatogenic defects\",\n        \"Mechanism by which WDR64 participates in intramanchette transport is entirely unresolved\",\n        \"No structural or biochemical characterization of WDR64\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"ODF1\"],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"WDR64 is a testis-specific WD40 repeat protein containing two β-propellers that dynamically localizes during spermiogenesis, associating with the manchette in elongating spermatids (steps 8–14) and subsequently relocating to the flagellum (steps 15–16) and the midpiece of mature spermatozoa [PMID:39386799]. WDR64 forms a complex with the outer dense fiber protein ODF1, co-localizing at both the manchette during nuclear shaping and the midpiece of mature sperm, implicating it in manchette-mediated nuclear remodeling and flagellar assembly [PMID:39386799].\",\n  \"teleology\": [\n    {\n      \"year\": 2024,\n      \"claim\": \"The first characterization of WDR64 established it as a testis-specific WD40 protein with a dynamic localization pattern during spermiogenesis—moving from the manchette to the flagellum—and identified ODF1 as a physical binding partner, linking WDR64 to nuclear shaping and flagellar assembly.\",\n      \"evidence\": \"Immunofluorescence, RT-PCR, Western blot, and IP-MS in mouse testis and spermatozoa (single lab study)\",\n      \"pmids\": [\"39386799\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No loss-of-function (knockout or knockdown) data demonstrating a requirement for WDR64 in spermiogenesis or fertility\",\n        \"WDR64–ODF1 interaction identified by IP-MS without reciprocal Co-IP or in vitro reconstitution\",\n        \"Structural basis for the two β-propeller domains and their individual contributions to localization or interaction is unknown\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Whether WDR64 is functionally required for sperm morphogenesis and male fertility remains untested, and the molecular mechanism by which the WDR64–ODF1 complex contributes to manchette-mediated nuclear shaping or flagellar midpiece assembly is unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No genetic studies in animal models or human cohorts linking WDR64 mutations to infertility or spermiogenesis defects\",\n        \"Downstream effectors or cargo transported by the WDR64–ODF1 complex are unidentified\",\n        \"Regulatory mechanisms controlling WDR64 expression timing and dynamic relocalization are unknown\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"ODF1\"],\n    \"other_free_text\": []\n  }\n}\n```"}